Importance of oil overlay for production of porcine embryos in vitro

Technologies to edit the zygote genome have revolutionized biomedical research not only for the creation of animal models for the study of human disease but also for the generation of functional human cells and tissues through interspecies blastocyst complementation technology. The pig is the ideal species for these purposes due to its great similarity in anatomy and physiology to humans. Emerging biotechnologies require the use of oocytes and/or embryos of good quality, which might be obtained using in vitro production (IVP) techniques. However, the current porcine embryo IVP systems are still suboptimal and result in low monospermic fertilization and blastocyst formation rates and poor embryo quality. During recent years, intensive investigations have been performed to evaluate the influence of specific compounds on gametes and embryos and to avoid the use of undefined supplements (serum and serum derivate) in the incubation media. However, little consideration has been given to the use of the mineral oil (MO) to overlay incubation droplets, which, albeit being a routine component of the IVP systems, is a totally undefined and thus problematic product for the safety of gametes and embryos. In this review, we provide an overview on the advantages and disadvantages of using MO to cover the incubation media. We also review one important concern in IVP laboratories: the use of oils containing undetected contamination. Finally, we discuss the effects of different types of oils on the in vitro embryo production outcomes and the transfer of compounds from oil into the culture media.     

Chrysanthemum cultivation in expanded clay. I. Effect of the nitrogen‐phosphorus‐potassium ratio in the nutrient solution

The hydroponic technique using expanded clay for the production of chrysanthemum was studied for possible use by Brazilian flower producers. Eight varying nutrient solution nitrogen (N):phosphorus (P):potassium (K) ratio treatments in a randomized block configuration with four replications were used. Six of the treatments supplied the same solution over the entire experimental period. The N:P:K ratios in the nutrient solution compared were: 1.0:0.3:1.0, 1.0:0.3:1.5, 1.0:0.3:2.0, 1.0:0.3:2.5, 1.5:0.3:1.0, and 2.0:0.3:1.0. The other two treatments consisted of N:P:K ratios of 2.0:0.3:1.0 during the juvenile stage and 1.0:0.3:1.5 or 1.0:0.3:2.0 ratios during the reproductive stage. There were two or three daily irrigations depending on plant need. The electrical conductivity (EC) and pH of the nutrient solution were checked once each day and the nutrient solution was chanced when 50% depleted. The plants were crown to the two‐stem stage under 50 long and 4.0 short days. Harvest was made 115 days after plant establishment. There were not expressive differences in plant characteristics among the treatments. The 1.0:0.3:2.5 N:P:K ratio gave the highest flower numbers per stem, stem length, and fresh and dry weights per plant.     

Macronutrient Accumulation in Coffee Fruits at Brazilian Zona Da Mata Conditions

Macronutrient accumulation in fruits and concentrations of these nutrients in leaves of fruit-bearing branches of arabica coffee were studied between anthesis and maturation. The experiment was carried out in a 2.0 × 1.0 m spacing of an arabica coffee orchard, collecting fruits and leaves of 18 randomized parcels at 12 sampling times, using a split-plot design. The largest accumulation rates of dry matter (DM), nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), and sulfur (S) were observed at the rapid expansion fruit stage, among 79 to 85 DAA (Days after the anthesis). Compared to the other nutrients, Ca and Mg accumulation occurred quickly in the 1st suspended growth stadium.     

URANIUM INDUCED EFFECTS ON DEVELOPMENT AND MINERAL NUTRITION OF ARABIDOPSIS THALIANA

This study aimed to investigate effects on growth and development and alterations in the nutrient profiles for Arabidopsis thaliana seedlings following uranium exposure. Seventeen-day-old Arabidopsis thaliana seedlings, grown in hydroponics, were exposed to 0, 0.1, 1, 10 and 100 μM uranium for one, three and seven days. Fresh weight of leaves and roots decreased after exposure to 100 μM uranium for one and three days and after exposure to 1, 10 and 100 μM uranium for seven days. Anthocyanous-colored leaves and stunted yellow roots were observed after exposure to 100 μM uranium. The uranium content of the roots highly increased but the root-to-shoot transfer of uranium was limited. Uranium exposure resulted in disturbed macro- and micronutrient profiles, especially following exposure to 100 μM. This study indicates that elevated uranium concentrations can cause important morphological and physiological effects and disturb the nutrient profiles in Arabidopsis thaliana seedlings.     

Index for potato nitrogen status diagnosis in a three-phase hydroponic system

Abstract

Up to now, there has not been any research that has tried to establish nitrogen (N) status diagnosis in basic seed potatoes, propagated by sprout in a three-phase hydroponic system. The objective of this study was to develop a potential index to be used in the diagnosis of N status in potato, using a three-phase hydroponic system, with the potato cultivar Agata. The treatments consisted of four N concentrations: 0, 7.8, 13 and 26?mmol L^?1. The experiment was conducted in a randomized block design with 12 replications in a non-acclimatized greenhouse. It was evaluated in a destructive and non-destructive way, and in real-time or space–time, 18 biometric characteristics of the reference leaf and of plant organs besides other variables related to the fourth leaf green color. Among the non-destructive characteristics measured, length, and diameter of the internodes and thickness of the fourth leaf showed the highest correlation coefficient (p?相似文献   

Occurrence of blossom‐end rot in tomato as a function of calcium dose in the nutrient solution and air relative humidity

The effects of different calcium (Ca) concentrations in the nutrient solution and of air relative humidity (RH) on the Ca levels and on the incidence of blossom‐end rot in tomato fruit cv. Jumbo were studied. The experiment was conducted in a greenhouse under hydroponic conditions using a modified Hoagland solution containing different Ca concentrations (0.2, 2.5, 5.0, 10.0, 15.0, and 20.0 mm L^‐1) which represented the different treatments. The trial was conducted in a fully randomized design with three replications, with Ca doses representing the plots and environments with high and low RH the subplots. The fruits of the second and third clusters were used, with two fruits per cluster, one maintained at high (90±5%) and the other at low (40±5%) RH. The fruits were picked after full ripening. A greater Ca accumulation was observed in fruits submitted to low RH with this accumulation occurring at all Ca levels in the solution. In the treatment with 100 mg Ca L^‐1, the plants died and did not reach fructification. The fruits of plants treated with 100 mg Ca L^‐1 and submitted to low RH had blossom‐end rot although their Ca levels were higher than those detected in the fruits of this treatment submitted to high RH. At this Ca dose, the seeds were small, malformed, and black. Our results show that fruit kept at low RH had higher Ca accumulation although the excessive water loss from tissues may lead to blossom‐end rot when low Ca doses are supplied to the plants.     

Specific detection by PCR of Streptococcus agalactiae in milk.

The aim of this study was to develop a simple and specific method for direct detection of Streptococcus agalactiae from cow's milk. The method was based on polymerase chain reaction (PCR) using species-specific and universal primers derived from the 16S rRNA gene. The amplification product was verified by restriction endonuclease digest and sequencing. Specific identification was proven on a collection of 147 S. agalactiae isolates of bovine and human origin. In addition, 17 strains belonging to different bacterial species that potentially can be found in milk samples also tested negative. The PCR developed was used for direct detection of S. agalactiae in milk, using for the first time with gram-positive bacteria the nucleic acid-binding properties of diatomaceous earth. The test, which has high specificity, high sensitivity (100 cfu/mL), and can be carried out in less than 24 h, represents an innovative diagnostic tool for the detection of S. agalactiae in milk.     

Ovine rumen papillae biopsy via oral endoscopy; a rapid and repeatable method for serial sampling

AIMS: To explore and validate the utility of rumen endoscopy for collection of rumen papillae for gene expression measurement.

METHODS: Four adult Coopworth ewes were fasted for either 4 or 24 hours. Animals were sedated, placed in a dorsally recumbent position at 45 degrees with the head upright, and an endoscope inserted via a tube inserted into the mouth. Biopsies of rumen papillae were taken from the ventral surface of the rumen atrium under visual guidance. Two biopsies were collected from one of the animals that had been fasted for 4 hours, and three from one of the animals that had been fasted for 24 hours. Video of the rumen atrium and reticulum was also collected. The animals recovered uneventfully. Biopsies were subsequently used for extraction and sequencing of mRNA.

RESULTS: The ventral surface of the rumen atrium was accessible after 4 hours off pasture, but a larger region was accessible after 24 hours of fasting. Sedation allowed access for endoscope use for around 5 to 10 minutes after which increased saliva flow was noted. Rumen papillae biopsies were easily collected, with samples from a variety of sites collected in the ～10 minute time window. High quality RNA was obtained for stranded mRNA sequencing. Of the resulting reads, 69–70% mapped uniquely to version 3.1 of the ovine genome, and 48–49% to a known gene. The rumen mRNA profiles were consistent with a previously reported study.

CONCLUSIONS: This method for obtaining rumenal tissue was found to be rapid and resulted in no apparent short or long term effects on the animal. High quality RNA was successfully extracted and amplified from the rumen papillae biopsies, indicating that this technique could be used for future gene expression studies. The use of rumen endoscopy could be extended to collection of a variety of rumen and reticulum anatomical measurements and deposition and retrieval of small sensors from the rumen. Rumen endoscopy offers an attractive and cost effective approach to repeated rumen biopsies compared with serial slaughter or use of cannulated animals.